PHARMACOLOGY OF THE CORNEAL ENDOTHELIUM

内皮细胞是维持角膜透明度的关键。使用 随着年龄的增长，人角膜内皮细胞失去分裂能力。自.以来 由于正常磨损而丢失的细胞没有替换的数量 组成单层的细胞随着时间的推移而减少。在老年人中， 疾病、创伤或眼内手术带来的压力会进一步损害 这种组织，导致它失去了保持完好无损的能力 单层，导致水肿，有时还需要角膜 移植。我们的长期目标是开发药理作用 治疗与年龄和疾病相关的角膜内皮变化。 要做到这一点，我们必须发现角膜内皮细胞如何对 通过断开与其邻居的接触而导致单层的缺陷 作为个体迁徙以重新定居该地区，和/或集体迁徙 放大和扁平，导致细胞的连续单层移动 或者扩散到缺陷区。使用模拟组织培养模型 成人角膜内皮的行为，我们发现 角膜内皮迁移和扩散在药物上是可分离的 运动的形式。我们将使用这些模型来：1)描述 区别迁移细胞和扩散细胞的形态特征，2) 确定细胞外基质成分是否可能是内源性的 迁移或传播的信号，3)识别细胞内 调节细胞迁移或扩散信号的途径，4) 确定在基因表达和蛋白质中发生了哪些修改 合成、磷酸化和亚细胞组织以产生 迁移或传播，以及5)确定特定的药理制剂 刺激伤口修复。将在这些方面使用的技术 研究包括扫描电子显微镜，数字化形态测量，一 以及双向凝胶电泳法，代谢标记法， 免疫细胞化学定位，Northern印迹分析，体外 翻译和原位杂交。使用这些工具获取的信息 方法应该为设计药物治疗提供基础，这将 保护人类角膜内皮功能，刺激，也许， 促进内皮细胞伤口愈合，减少应激对皮肤的影响 老化的内皮细胞。我们的研究还将建立角膜 内皮细胞作为单层组织创伤修复研究的模型 在眼睛和其他器官里。

The endothelium is critical for maintaining corneal transparency. With age, human corneal endothelial cells lose their ability to divide. Since there is no replacement of cells lost due to normal attrition the number of cells comprising the monolayer decreases over time. In older persons, the stress of disease, trauma, or intraocular surgery can further compromise this tissue, causing it to lose its ability to maintain an intact monolayer, producing edema and, sometimes, the need for corneal transplantation. Our long-term goal is to develop pharmacological treatment for age- and disease-related changes in the corneal endothelium. To do this, we must discover how corneal endothelial cells respond to a defect in the monolayer by breaking contact with their neighbors and migrating as individuals to repopulate the area, and/or by collectively enlarging and flattening, causing the continuous monolayer of cells to move or spread into the defect area. Using tissue culture models which mimic the behavior of adult human corneal endothelium, we have discovered that corneal endothelial migration and spreading are pharmacologically separable forms of movement. We will use these models to: 1) characterize the morphologic features which distinguish migrating from spreading cells, 2) determine whether extracellular matrix components might be endogenous signals for either migration or spreading, 3) identify the intracellular pathway which mediates the signal for cell migration or for spreading, 4) determine what modifications occur in gene expression and in protein synthesis, phosphorylation and subcellular organization to produce either migration or spreading, and 5) identify specific pharmacological agents which stimulate wound repair. Techniques that will be used in these studies include scanning electron microscopy, digitized morphometry, one and two-dimensional gel electrophoresis, metabolic labeling, immunocytochemical localization, Northern blot analysis, in vitro translation and in situ hybridization. Information obtained using these methods should provide a basis for designing drug treatments which will preserve human corneal endothelial function, stimulate and, perhaps, accelerate endothelial wound healing, and reduce the effects of stress on the aging endothelium. Our studies will also establish the corneal endothelium as a model for the study of wound repair in monolayer tissues in the eye and in other organs.